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Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum, and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction
Lu, Y; Rong, CZ; Zhao, JY; Lao, XJ; Xie, L; Li, S; Qin, X.
  • Lu, Y; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
  • Rong, CZ; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
  • Zhao, JY; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
  • Lao, XJ; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
  • Xie, L; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
  • Li, S; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
  • Qin, X; Guangxi Medical University. Department of Clinical Laboratory. Nanning. CN
Braz. j. med. biol. res ; 49(10): e5303, 2016. tab, graf
Article in English | LILACS | ID: lil-792526
ABSTRACT
The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period.
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Full text: Available Index: LILACS (Americas) Main subject: Specimen Handling / DNA, Bacterial / Chlamydia trachomatis / Ureaplasma urealyticum / Real-Time Polymerase Chain Reaction / Neisseria gonorrhoeae Type of study: Diagnostic study Limits: Adult / Female / Humans / Male Language: English Journal: Braz. j. med. biol. res Journal subject: Biology / Medicine Year: 2016 Type: Article / Project document Affiliation country: China Institution/Affiliation country: Guangxi Medical University/CN

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Full text: Available Index: LILACS (Americas) Main subject: Specimen Handling / DNA, Bacterial / Chlamydia trachomatis / Ureaplasma urealyticum / Real-Time Polymerase Chain Reaction / Neisseria gonorrhoeae Type of study: Diagnostic study Limits: Adult / Female / Humans / Male Language: English Journal: Braz. j. med. biol. res Journal subject: Biology / Medicine Year: 2016 Type: Article / Project document Affiliation country: China Institution/Affiliation country: Guangxi Medical University/CN